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Updated: Apr 26, 2026

A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors

A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors

Published on: April 29, 2022

In situ sample processing approach (iSPA) for comprehensive quantitative phosphoproteome analysis.

Junfeng Huang¹, Hongqiang Qin, Jing Dong

¹CAS Key Lab of Separation Sciences for Analytical Chemistry National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , Dalian 116023, China.

Journal of Proteome Research

|July 30, 2014

Summary

This summary is machine-generated.

A new in situ sample processing approach (iSPA) simplifies quantitative phosphoproteome analysis. This method reduces steps, minimizes sample loss, and improves accuracy for analyzing phosphorylation sites in complex biological samples like liver tissue.

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Published on: July 26, 2019

Area of Science:

Biochemistry
Proteomics
Analytical Chemistry

Background:

Current phosphoproteome analysis methods are complex and time-consuming.
Efficient sample preparation is crucial for accurate quantitative analysis.

Purpose of the Study:

To develop a streamlined in situ sample processing approach (iSPA) for quantitative phosphoproteome analysis.
To improve the efficiency, sensitivity, and accuracy of phosphoproteome analysis.

Main Methods:

Utilized macroporous Ti(IV)-IMAC microspheres for in situ enrichment, digestion, and labeling.
Integrated multiple sample preparation steps sequentially on the microspheres.
Avoided traditional desalting and buffer exchange steps.

Main Results:

Achieved seamless sample processing, reducing sample loss and contamination.
Demonstrated enhanced sensitivity and accuracy in quantitative phosphoproteome analysis.
Successfully applied iSPA to differential phosphoproteome analysis of human liver tissues, quantifying 8548 phosphorylation sites.

Conclusions:

The iSPA strategy offers a facile and efficient method for high-quality quantitative phosphoproteome analysis.
This approach is suitable for analyzing complex biological samples and identifying differential phosphorylation in disease contexts like hepatocellular carcinoma (HCC).